Method and kit for quantitatively measuring beta-glucuronidase in multiple samples by enzyme-linked immunosorbent assay (ELIASA) instrument

Abstract

The invention relates to a method and a kit for quantitatively measuring the beta-glucuronidase in multiple samples by an enzyme-linked immunosorbent assay (ELIASA) instrument. The kit internally comprises beta-glucuronidase standard substance, working solution, phenolphthalein glucuronic acid reaction liquid, termination reaction reagent, a pH meter and the like. The method and the kit have the advantages that the concentration of the beta-glucuronidase in the multiple samples can be detected at the same time, and shorter detection time is consumed; the sensitivity is high, and the detection limit is 39.0625U / mL; the method is wide in linear range, the detection concentration is within the range of 39.0625-12500U / mL, and the three order of magnitudes are spanned; the accuracy is high; less reagent and fewer samples are used; the kit is convenient to operate; the samples can be fed once without being fed for a plurality of times; the cleaning step is omitted, so that the operation steps are reduced, cross contamination is prevented, and the result is guaranteed to be credible; the detected samples can be well stored so as to be repeatedly detected.

Description

Technical field: [0001] The invention belongs to the field of biochemical analysis, in particular to a method and a detection kit for quantitatively and simultaneously detecting β-glucuronidase in a plurality of samples with a microplate reader. Background technique: [0002] β-glucuronidase is a lysosomal enzyme that promotes the hydrolysis of the β-D-glycosidic bonds of many glucosides in the body. [0003] In the current technical method, the method for detecting β-glucuronidase in feces is limited to ultraviolet spectrophotometer and automatic biochemical analyzer. [0004] Both methods share the following disadvantages: [0005] 1. A large amount of reagents are used [0006] 2. Need more samples [0007] In addition, ultraviolet spectrophotometers and automatic biochemical analyzers require a large amount of samples. If the sample size is not enough, the detection cannot be performed. [0008] 3. It takes a long time [0009] When multiple samples need to be detec...

AI Technical Summary

Problems solved by technology

[0005] 1. A large amount of reagents are used

[0006] 2. A large number of samples are required

[0007] In addition, ultraviolet spectrophotometer and automatic biochemical analyzer require a large amount of samples. If the sample size is not enough, the detection cannot be carried out.

[0008] 3. It takes a long time

[0011] In addition to the inaccurate results caused by the long time-consuming of the automatic biochemical instrument and the UV spectrophotometer, there may also be the possibility of cross-contamination of samples or standards

[0012] 4. Cleaning steps required

[0014] 5. Inaccurate results

Since the absorbance of the standard curve (usually five points) is not read at the same time, the time for detecting the enzyme activity is not synchronized, and the time extension will reduce the detected enzyme activity, so the standard curve obtained will be different, and is an unavoidable systematic error

In addition, the sampling and testing of each sample is a manual operation, and there will be operational errors

[0016] The detection of samples by the automatic biochemical instrument is better than that of the ultraviolet spectrophotometer, mainly in the accuracy of adding samples. However, the detection time of each sample has not been shortened, and the operation of the automatic biochemical instrument still has a An indispensable step is to clean the injection needle, so that if the cleaning is not thorough, it will cause cross-contamination, which will lead to unreliable results

[0017] 6. It is not conducive to the preservation of original samples

[0018] The amount of sample required for UV spectrophotometer detection is large (even with the smallest amount of cuvette, due to the limitation of the method, the sample amount is 1mL). After the detection, because multiple samples use the same group (2~ 3) cuvettes, so compared with the original sample, the sample after recovery and detection has the possibility of contamination in theory and actual operation, which is not conducive to sample preservation

[0019] Although the sample volume required for the detection of samples by the automatic biochemical analyzer is less than that of the ultraviolet spectrophotometer (200 μL to 500 μL), the samples after each detection cannot be saved due to the detection method

Images